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1.
Clin Lab ; 70(5)2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38747929

RESUMO

BACKGROUND: Female vulvovaginitis was one of the most common gynecological diseases. It had a great negative impact on their work and quality of life. This retrospective study evaluated the clinical and laboratory data of patients with vulvovaginitis in Hangzhou, China. To analyze the clinical situation, species distribution and antibiotic resistance of pathogenic fungi and bacteria in 626 cases of vulvovaginitis in Hangzhou. Microorganism culture, identification, and antibiotic susceptibility testing were conducted. The study aimed to provide a theoretical value for an effective treatment of vulvovaginitis. METHODS: In total, 626 outpatients and inpatients diagnosed with vulvovaginitis were selected from January 2018 to January 2023. Data of all the patients were collected from the hospital's electronic medical records. Vaginal secretion was collected for testing and SPSS 25.0 software was used to perform statistical analysis. RESULTS: A total of 626 strains of fungi, Gram-positive, and -negative bacteria were detected. Clinical situations of patients infected with the top five pathogenic fungi and bacteria were analyzed. Pathogenic fungi and bacteria were slightly different in each age group and in each onset time group. The results of antibiotic susceptibility testing showed that the resistance rates of itraconazole and fluconazole were high and Gram- negative and -positive bacteria were multidrug resistant. Gram-negative bacteria were more sensitive to carbenicillins and compound antibiotics, while Gram-positive bacteria were sensitive to rifampicin and daptomycin. MRSA and non vancomycin-resistant strains were detected. CONCLUSIONS: Fungi and bacteria were usually detected as pathogenes in patients with vulvovaginitis in Hangzhou. Some factors, such as age and onset time, often affected the incidence. Pathogenic fungi and bacteria were resistant to some common antibiotics, and clinical treatments should be carried out in a timely and reasonable manner according to the results of antibiotic susceptibility testing.


Assuntos
Fungos , Testes de Sensibilidade Microbiana , Vulvovaginite , Humanos , Feminino , China/epidemiologia , Adulto , Vulvovaginite/microbiologia , Vulvovaginite/tratamento farmacológico , Vulvovaginite/epidemiologia , Vulvovaginite/diagnóstico , Estudos Retrospectivos , Fungos/efeitos dos fármacos , Fungos/isolamento & purificação , Fungos/classificação , Pessoa de Meia-Idade , Adulto Jovem , Adolescente , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Bactérias/classificação , Farmacorresistência Fúngica , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Farmacorresistência Bacteriana , Idoso
2.
Sci Rep ; 14(1): 10544, 2024 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-38719860

RESUMO

The increasing amount of weeds surviving herbicide represents a very serious problem for crop management. The interaction between microbial community of soil and herbicide resistance, along with the potential evolutive consequences, are still poorly known and need to be investigated to better understand the impact on agricultural management. In our study, we analyzed the microbial composition of soils in 32 farms, located in the Northern Italy rice-growing area (Lombardy) with the aim to evaluate the relationship between the microbial composition and the incidence of resistance to acetolactate synthase (ALS) and acetyl-CoA carboxylase (ACCase) inhibiting herbicides in Echinochloa species. We observed that the coverage of weeds survived herbicide treatment was higher than 60% in paddy fields with a low microbial biodiversity and less than 5% in those with a high microbial biodiversity. Fungal communities showed a greater reduction in richness than Bacteria. In soils with a reduced microbial diversity, a significant increase of some bacterial and fungal orders (i.e. Lactobacillales, Malasseziales and Diaporthales) was observed. Interestingly, we identified two different microbial profiles linked to the two conditions: high incidence of herbicide resistance (H-HeR) and low incidence of herbicide resistance (L-HeR). Overall, the results we obtained allow us to make hypotheses on the greater or lesser probability of herbicide resistance occurrence based on the composition of the soil microbiome and especially on the degree of biodiversity of the microbial communities.


Assuntos
Acetolactato Sintase , Acetil-CoA Carboxilase , Echinochloa , Resistência a Herbicidas , Herbicidas , Microbiologia do Solo , Itália/epidemiologia , Herbicidas/farmacologia , Acetolactato Sintase/antagonistas & inibidores , Acetolactato Sintase/genética , Echinochloa/efeitos dos fármacos , Acetil-CoA Carboxilase/genética , Acetil-CoA Carboxilase/antagonistas & inibidores , Plantas Daninhas/efeitos dos fármacos , Microbiota/efeitos dos fármacos , Biodiversidade , Bactérias/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/classificação , Solo/química , Fungos/efeitos dos fármacos , Fungos/isolamento & purificação , Fungos/genética
3.
Curr Microbiol ; 81(6): 160, 2024 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-38695903

RESUMO

Salt stress can adversely affect plant seed germination, growth and development, and eventually lead to slow growth and even death of plants. The purpose of this study was to investigate the effects of different concentrations of NaCl and Na2SO4 stress on the physicochemical properties, enzyme activities, rhizosphere microbial community and seven active components (L-phenylalanine, Protocatechuic acid, Eleutheroside B, Chlorogenic acid, Caffeic acid, Eleutheroside E, Isofraxidin) of Acanthopanax senticosus rhizosphere soil. Statistical analysis was used to explore the correlation between the rhizosphere ecological factors of Acanthopanax senticosus and its active components. Compared with Acanthopanax senticosus under NaCl stress, Na2SO4 generally had a greater effect on Acanthopanax senticosus, which reduced the richness of fungi in rhizosphere soil and adversely affected the content of multiple active components. Pearson analysis showed that pH, organic matter, ammonium nitrogen, available phosphorus, available potassium, catalase and urease were significantly correlated with active components such as Caffeic acid and Isofraxidin. There were 11 known bacterial genera, 12 unknown bacterial genera, 9 known fungal genera and 1 unknown fungal genus significantly associated with the active ingredient. Salt stress had great changes in the physicochemical properties, enzyme activities and microorganisms of the rhizosphere soil of Acanthopanax senticosus. In conclusion, different types and concentrations of salts had different effects on Acanthopanax senticosus, and the active components of Acanthopanax senticosus were regulated by rhizosphere soil ecological factors.


Assuntos
Bactérias , Eleutherococcus , Fungos , Rizosfera , Estresse Salino , Microbiologia do Solo , Bactérias/classificação , Bactérias/genética , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Bactérias/metabolismo , Fungos/classificação , Fungos/efeitos dos fármacos , Fungos/genética , Fungos/isolamento & purificação , Eleutherococcus/metabolismo , Microbiota/efeitos dos fármacos , Solo/química , Cloreto de Sódio/metabolismo , Raízes de Plantas/microbiologia
4.
Environ Microbiol Rep ; 16(3): e13213, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38738810

RESUMO

Since a significant proportion of plant matter is consumed by herbivores, a necessary adaptation for many phyllosphere microbes could be to survive through the guts of herbivores. While many studies explore the gut microbiome of herbivores by surveying the microbiome in their frass, few studies compare the phyllosphere microbiome to the gut microbiome of herbivores. High-throughput metabarcode sequencing was used to track the fungal community from milkweed (Asclepias spp.) leaves to monarch caterpillar frass. The most commonly identified fungal taxa that dominated the caterpillar frass after the consumption of leaves were yeasts, mostly belonging to the Basidiomycota phylum. While most fungal communities underwent significant bottlenecks and some yeast taxa increased in relative abundance, a consistent directional change in community structure was not identified from leaf to caterpillar frass. These results suggest that some phyllosphere fungi, especially diverse yeasts, can survive herbivory, but whether herbivory is a key stage of their life cycle remains uncertain. For exploring phyllosphere fungi and the potential coprophilous lifestyles of endophytic and epiphytic fungi, methods that target yeast and Basidiomycota fungi are recommended.


Assuntos
Asclepias , Fungos , Herbivoria , Folhas de Planta , Animais , Folhas de Planta/microbiologia , Asclepias/microbiologia , Fungos/classificação , Fungos/genética , Fungos/isolamento & purificação , Fungos/fisiologia , Leveduras/classificação , Leveduras/isolamento & purificação , Leveduras/genética , Micobioma , Basidiomycota/classificação , Basidiomycota/genética , Basidiomycota/fisiologia , Basidiomycota/isolamento & purificação , Microbioma Gastrointestinal , Larva/microbiologia , Mariposas/microbiologia
5.
PLoS One ; 19(5): e0301605, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38739592

RESUMO

Oxygen minimum zones (OMZ) represent ~8% of the ocean, with the Pacific as the largest and top expanding area. These regions influence marine ecosystems, promoting anaerobic microbial communities. Nevertheless, only a fraction of microbial diversity has been studied, with fungi being the less explored component. So, herein we analyzed fungal diversity patterns in surface and subsurface sediments along a bathymetric transect using metabarcoding of the ITS1 region in the OMZ of the Mexican Pacific off Mazatlán. We identified 353 amplicon sequence variants (ASV), within the Ascomycota, Basidiomycota, and Rozellomycota. Spatial patterns evidenced higher alpha diversity in nearshore and subsurface subsamples, probably due to temporal fluctuations in organic matter inputs. Small-scale heterogeneity characterized the community with the majority of ASV (269 ASV) occurring in a single subsample, hinting at the influence of local biogeochemical conditions. This baseline data evidenced a remarkable fungal diversity presenting high variation along a bathymetric and vertical transects.


Assuntos
Biodiversidade , Código de Barras de DNA Taxonômico , Fungos , Sedimentos Geológicos , Oxigênio , Sedimentos Geológicos/microbiologia , Oxigênio/metabolismo , Oxigênio/análise , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Oceano Pacífico , Filogenia
6.
EBioMedicine ; 103: 105137, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38703606

RESUMO

BACKGROUND: Coronary artery disease (CAD) is a prevalent cardiovascular condition, and numerous studies have linked gut bacterial imbalance to CAD. However, the relationship of gut fungi, another essential component of the intestinal microbiota, with CAD remains poorly understood. METHODS: In this cross-sectional study, we analyzed fecal samples from 132 participants, split into 31 healthy controls and 101 CAD patients, further categorized into stable CAD (38), unstable angina (41), and acute myocardial infarction (22) groups. We conducted internal transcribed spacer 1 (ITS1) and 16S sequencing to examine gut fungal and bacterial communities. FINDINGS: Based on ITS1 analyses, Ascomycota and Basidiomycota were the dominant fungal phyla in all the groups. The α diversity of gut mycobiome remained unaltered among the control group and CAD subgroups; however, the structure and composition of the mycobiota differed significantly with the progression of CAD. The abundances of 15 taxa gradually changed with the occurrence and progression of the disease and were significantly correlated with major CAD risk factor indicators. The mycobiome changes were closely linked to gut microbiome dysbiosis in patients with CAD. Furthermore, disease classifiers based on gut fungi effectively identified subgroups with different degrees of CAD. Finally, the FUNGuild analysis further categorized these fungi into distinct ecological guilds. INTERPRETATION: In conclusion, the structure and composition of the gut fungal community differed from healthy controls to various subtypes of CAD, revealing key fungi taxa alterations linked to the onset and progression of CAD. Our study highlights the potential role of gut fungi in CAD and may facilitate the development of novel biomarkers and therapeutic targets for CAD. FUNDING: This work was supported by the grants from the National Natural Science Foundation of China (No. 82170302, 92168117, 82370432), National clinical key specialty construction project- Cardiovascular Surgery, the Reform and Development Program of Beijing Institute of Respiratory Medicine (No. Ggyfz202417, Ggyfz202308), the Beijing Natural Science Foundation (No. 7222068); and the Clinical Research Incubation Program of Beijing Chaoyang Hospital Affiliated to Capital Medical University (No. CYFH202209).


Assuntos
Doença da Artéria Coronariana , Microbioma Gastrointestinal , Micobioma , Humanos , Doença da Artéria Coronariana/microbiologia , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Estudos Transversais , Fezes/microbiologia , Metagenômica/métodos , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Índice de Gravidade de Doença , Disbiose/microbiologia , Estudos de Casos e Controles , RNA Ribossômico 16S/genética , Adulto
7.
Microb Ecol ; 87(1): 66, 2024 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-38700528

RESUMO

Despite the importance of wood-inhabiting fungi on nutrient cycling and ecosystem functions, their ecology, especially related to their community assembly, is still highly unexplored. In this study, we analyzed the wood-inhabiting fungal richness, community composition, and phylogenetics using PacBio sequencing. Opposite to what has been expected that deterministic processes especially environmental filtering through wood-physicochemical properties controls the community assembly of wood-inhabiting fungal communities, here we showed that both deterministic and stochastic processes can highly contribute to the community assembly processes of wood-inhabiting fungi in this tropical forest. We demonstrated that the dynamics of stochastic and deterministic processes varied with wood decomposition stages. The initial stage was mainly governed by a deterministic process (homogenous selection), whereas the early and later decomposition stages were governed by the stochastic processes (ecological drift). Deterministic processes were highly contributed by wood physicochemical properties (especially macronutrients and hemicellulose) rather than soil physicochemical factors. We elucidated that fine-scale fungal-fungal interactions, especially the network topology, modularity, and keystone taxa of wood-inhabiting fungal communities, strongly differed in an initial and decomposing deadwood. This current study contributes to a better understanding of the ecological processes of wood-inhabiting fungi in tropical regions where the knowledge of wood-inhabiting fungi is highly limited.


Assuntos
Florestas , Fungos , Micobioma , Madeira , Madeira/microbiologia , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Clima Tropical , Filogenia , Sequenciamento de Nucleotídeos em Larga Escala , Biodiversidade
8.
Food Res Int ; 183: 114196, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38760131

RESUMO

Baijiu production has relied on natural inoculated Qu as a starter culture, causing the unstable microbiota of fermentation grains, which resulted in inconsistent product quality across batches. Therefore, revealing the core microbes and constructing a synthetic microbiota during the fermentation process was extremely important for stabilizing product quality. In this study, the succession of the microbial community was analyzed by high-throughput sequencing technology, and ten core microbes of Xiaoqu light-aroma Baijiu were obtained by mathematical statistics, including Acetobacter, Bacillus, Lactobacillus, Weissella, Pichia,Rhizopus, Wickerhamomyces, Issatchenkia, Saccharomyces, and Kazachstania. Model verification showed that the core microbiota significantly affected the composition of non-core microbiota (P < 0.01) and key flavor-producing enzymes (R > 0.8, P < 0.01), thus significantly affecting the flavor of base Baijiu. Simulated fermentation validated that the core microbiota can reproduce the fermentation process and quality of Xiaoqu light-aroma Baijiu. The succession of bacteria was mainly regulated by acidity and ethanol, while the fungi, especially non-Saccharomyces cerevisiae, were mainly regulated by the initial dominant bacteria (Acetobacter, Bacillus, and Weissella). This study will play an important role in the transformation of Xiaoqu light-aroma Baijiu fermentation from natural fermentation to controlled fermentation and the identification of core microbes in other fermented foods.


Assuntos
Bactérias , Fermentação , Microbiologia de Alimentos , Microbiota , Bactérias/classificação , Bactérias/metabolismo , Bactérias/genética , Bactérias/isolamento & purificação , Fungos/genética , Fungos/classificação , Fungos/metabolismo , Fungos/isolamento & purificação , Bebidas Alcoólicas/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala , Paladar , Aromatizantes/metabolismo
9.
Microb Ecol ; 87(1): 72, 2024 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-38755460

RESUMO

Air pollution caused by tropospheric ozone contributes to the decline of forest ecosystems; for instance, sacred fir, Abies religiosa (Kunth) Schltdl. & Cham. forests in the peri-urban region of Mexico City. Individual trees within these forests exhibit variation in their response to ozone exposure, including the severity of visible symptoms in needles. Using RNA-Seq metatranscriptomic data and ITS2 metabarcoding, we investigated whether symptom variation correlates with the taxonomic and functional composition of fungal mycobiomes from needles collected in this highly polluted area in the surroundings of Mexico City. Our findings indicate that ozone-related symptoms do not significantly correlate with changes in the taxonomic composition of fungal mycobiomes. However, genes coding for 30 putative proteins were differentially expressed in the mycobiome of asymptomatic needles, including eight genes previously associated with resistance to oxidative stress. These results suggest that fungal communities likely play a role in mitigating the oxidative burst caused by tropospheric ozone in sacred fir. Our study illustrates the feasibility of using RNA-Seq data, accessible from global sequence repositories, for the characterization of fungal communities associated with plant tissues, including their gene expression.


Assuntos
Poluição do Ar , Fungos , Micobioma , Folhas de Planta , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Folhas de Planta/microbiologia , México , Poluição do Ar/efeitos adversos , Ozônio , Estresse Fisiológico , Cidades
10.
World J Microbiol Biotechnol ; 40(7): 205, 2024 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-38755302

RESUMO

Jojoba shrubs are wild plants cultivated in arid and semiarid lands and characterized by tolerance to drought, salinity, and high temperatures. Fungi associated with such plants may be attributed to the tolerance of host plants against biotic stress in addition to the promotion of plant growth. Previous studies showed the importance of jojoba as jojoba oil in the agricultural field; however, no prior study discussed the role of jojoba-associated fungi (JAF) in reflecting plant health and the possibility of using JAF in biocontrol. Here, the culture-independent and culture-dependent approaches were performed to study the diversity of the jojoba-associated fungi. Then, the cultivable fungi were evaluated for in-vitro antagonistic activity and in vitro plant growth promotion assays. The metagenome analysis revealed the existence of four fungal phyla: Ascomycota, Aphelidiomycota, Basidiomycota, and Mortierellomycota. The phylum Ascomycota was the most common and had the highest relative abundance in soil, root, branch, and fruit samples (59.7%, 50.7%, 49.8%, and 52.4%, respectively). Alternaria was the most abundant genus in aboveground tissues: branch (43.7%) and fruit (32.1%), while the genus Discosia had the highest abundance in the underground samples: soil (24%) and root (30.7%). For the culture-dependent method, a total of 14 fungi were isolated, identified, and screened for their chitinolytic and antagonist activity against three phytopathogenic fungi (Fusarium oxysporum, Alternaria alternata and Rhizoctonia solani) as well as their in vitro plant growth promotion (PGP) activity. Based on ITS sequence analysis, the selected potent isolates were identified as Aspergillus stellatusEJ-JFF3, Aspergillus flavus EJ-JFF4, Stilbocrea sp. EJ-JLF1, Fusarium solani EJ-JRF3, and Amesia atrobrunneaEJ-JSF4. The endophyte strain A. flavus EJ-JFF4 exhibited the highest chitinolytic activity (9 Enzyme Index) and antagonistic potential against Fusarium oxysporum, Alternaria alternata, and Rhizoctonia solani phytopathogens with inhibitory percentages of 72, 70, and 80 respectively. Also, A. flavus EJ-JFF4 had significant multiple PGP properties, including siderophore production (69.3%), phosphate solubilization (95.4 µg ml-1). The greatest production of Indol-3-Acetic Acid was belonged to A. atrobrunnea EJ-JSF4 (114.5 µg ml-1). The analysis of FUNGuild revealed the abundance of symbiotrophs over other trophic modes, and the guild of endophytes was commonly assigned in all samples. For the first time, this study uncovered fungal diversity associated with jojoba plants using a culture-independent approach and in-vitro assessed the roles of cultivable fungal strains in promoting plant growth and biocontrol. The present study indicated the significance of jojoba shrubs as a potential source of diverse fungi with high biocontrol and PGP activities.


Assuntos
Alternaria , Fungos , Microbiologia do Solo , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Alternaria/genética , Alternaria/crescimento & desenvolvimento , Metagenoma , Rhizoctonia/crescimento & desenvolvimento , Filogenia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Fusarium/genética , Fusarium/crescimento & desenvolvimento , Antibiose , Raízes de Plantas/microbiologia , Biodiversidade , Agentes de Controle Biológico , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/genética , Desenvolvimento Vegetal
11.
PLoS One ; 19(5): e0302462, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38753836

RESUMO

Fruit shape is an important character of watermelon. And the compositions of rhizospheric and endophytic microorganisms of watermelon with different fruit shape also remains unclear. To elucidate the biological mechanism of watermelon fruit shape formations, the rhizospheric and endophytic microbial community compositions between oval (OW) and circular watermelons (CW) were analyzed. The results showed that except of the rhizospheric bacterial richness (P < 0.05), the rhizospheric and endophytic microbial (bacterial and fungal) diversity were not statistically significant between OW and CW (P > 0.05). However, the endophytic microbial (bacterial and fungal) compositions were significantly different. Firstly, Bacillus, Rhodanobacter, Cupriavidus, Luteimonas, and Devosia were the unique soil dominant bacterial genera in rhizospheres of circular watermelon (CW); In contrast, Nocardioides, Ensifer, and Saccharomonospora were the special soil dominant bacterial genera in rhizospheres of oval watermelons (OW); Meanwhile, Cephalotrichum, Neocosmospora, Phialosimplex, and Papulaspora were the unique soil dominant fungal genera in rhizospheres of circular watermelon (CW); By contrast, Acremonium, Cladosporium, Cryptococcus_f__Tremellaceae, Sodiomyces, Microascus, Conocybe, Sporidiobolus, and Acremonium were the unique soil dominant fungal genera in rhizospheres of oval watermelons (OW). Additionally, Lechevalieria, Pseudorhodoferax, Pseudomonas, Massilia, Flavobacterium, Aeromicrobium, Stenotrophomonas, Pseudonocardia, Novosphingobium, Melittangium, and Herpetosiphon were the unique dominant endophytic bacterial genera in stems of CW; In contrast, Falsirhodobacter, Kocuria, and Kineosporia were the special dominant endophytic genera in stems of OW; Moreover, Lectera and Fusarium were the unique dominant endophytic fungal genera in stems of CW; By contrast, Cercospora only was the special dominant endophytic fungal genus in stems of OW. All above results suggested that watermelons with different fruit shapes exactly recruited various microorganisms in rhizospheres and stems. Meanwhile, the enrichments of the different rhizosphric and endophytic microorganisms could be speculated in relating to watermelon fruit shapes formation.


Assuntos
Bactérias , Citrullus , Endófitos , Frutas , Fungos , Rizosfera , Microbiologia do Solo , Citrullus/microbiologia , Endófitos/genética , Frutas/microbiologia , Bactérias/genética , Bactérias/classificação , Bactérias/isolamento & purificação , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Microbiota/genética
12.
Anal Chem ; 96(19): 7787-7796, 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38702857

RESUMO

Microorganism are ubiquitous and intimately connected with human health and disease management. The accurate and fast identification of pathogenic microorganisms is especially important for diagnosing infections. Herein, three tetraphenylethylene derivatives (S-TDs: TBN, TPN, and TPI) featuring different cationic groups, charge numbers, emission wavelengths, and hydrophobicities were successfully synthesized. Benefiting from distinct cell wall binding properties, S-TDs were collectively utilized to create a sensor array capable of imaging various microorganisms through their characteristic fluorescent signatures. Furthermore, the interaction mechanism between S-TDs and different microorganisms was explored by calculating the binding energy between S-TDs and cell membrane/wall constituents, including phospholipid bilayer and peptidoglycan. Using a combination of the fluorescence sensor array and a deep learning model of residual network (ResNet), readily differentiation of Gram-negative bacteria (G-), Gram-positive bacteria (G+), fungi, and their mixtures was achieved. Specifically, by extensive training of two ResNet models with large quantities of images data from 14 kinds of microorganism stained with S-TDs, identification of microorganism was achieved at high-level accuracy: over 92.8% for both Gram species and antibiotic-resistant species, with 90.35% accuracy for the detection of mixed microorganism in infected wound. This novel method provides a rapid and accurate method for microbial classification, potentially aiding in the diagnosis and treatment of infectious diseases.


Assuntos
Aprendizado Profundo , Humanos , Estilbenos/química , Bactérias Gram-Positivas/isolamento & purificação , Corantes Fluorescentes/química , Bactérias Gram-Negativas/isolamento & purificação , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/diagnóstico , Fungos/isolamento & purificação
13.
Sci Rep ; 14(1): 10899, 2024 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-38740854

RESUMO

In order to obtain the best mass spectrometry identification results for using the most appropriate methods in clinical practice, we explore the optimal pretreatment methods for different species and morphologies of filamentous fungi. 98 fungal strains were treated with formic acid sandwich method, dispersion method, extraction method, and other methods using a medium element mass spectrometer (EXS3000) as a platform. Each strain had three targets, and the identification rates and confidence differences under different pre-treatment methods were compared to evaluate the identification effects of these methods. The mass spectrometry identification rates of 98 filamentous fungi obtained after pre-treatment with formic acid sandwich method, dispersion method, and extraction method were 85.71%, 82.65%, and 75.51%, respectively. The identification rate of the formic acid sandwich method was significantly higher than the other two methods (P < 0 005) has the best identification ability and the obtained confidence is also higher than the other two methods. The use of formic acid sandwich method for mass spectrometry identification of filamentous fungi can achieve ideal identification results, which is suitable for mass spectrometry identification of filamentous fungi in conventional laboratories.


Assuntos
Fungos , Espectrometria de Massas , Fungos/isolamento & purificação , Fungos/classificação , Espectrometria de Massas/métodos , Formiatos/química , Formiatos/análise , Micoses/microbiologia , Micoses/diagnóstico , Humanos
14.
J Clin Microbiol ; 62(5): e0174923, 2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38624235

RESUMO

The timely identification of microbial pathogens is essential to guide targeted antimicrobial therapy and ultimately, successful treatment of an infection. However, the yield of standard microbiology testing (SMT) is directly related to the duration of antecedent antimicrobial therapy as SMT culture methods are dependent on the recovery of viable organisms, the fastidious nature of certain pathogens, and other pre-analytic factors. In the last decade, metagenomic next-generation sequencing (mNGS) has been successfully utilized as a diagnostic tool for various applications within the clinical laboratory. However, mNGS is resource, time, and labor-intensive-requiring extensive laborious preliminary benchwork, followed by complex bioinformatic analysis. We aimed to address these shortcomings by developing a largely Automated targeted Metagenomic next-generation sequencing (tmNGS) PipeLine for rapId inFectIous disEase Diagnosis (AMPLIFIED) to detect bacteria and fungi directly from clinical specimens. Therefore, AMPLIFIED may serve as an adjunctive approach to complement SMT. This tmNGS pipeline requires less than 1 hour of hands-on time before sequencing and less than 2 hours of total processing time, including bioinformatic analysis. We performed tmNGS on 50 clinical specimens with concomitant cultures to assess feasibility and performance in the hospital laboratory. Of the 50 specimens, 34 (68%) were from true clinical infections. Specimens from cases of true infection were more often tmNGS positive compared to those from the non-infected group (82.4% vs 43.8%, respectively, P = 0.0087). Overall, the clinical sensitivity of AMPLIFIED was 54.6% with 85.7% specificity, equating to 70.6% and 75% negative and positive predictive values, respectively. AMPLIFIED represents a rapid supplementary approach to SMT; the typical time from specimen receipt to identification of potential pathogens by AMPLIFIED is roughly 24 hours which is markedly faster than the days, weeks, and months required to recover bacterial, fungal, and mycobacterial pathogens by culture, respectively. IMPORTANCE: To our knowledge, this represents the first application of an automated sequencing and bioinformatics pipeline in an exclusively pediatric population. Next-generation sequencing is time-consuming, labor-intensive, and requires experienced personnel; perhaps contributing to hesitancy among clinical laboratories to adopt such a test. Here, we report a strong case for use by removing these barriers through near-total automation of our sequencing pipeline.


Assuntos
Bactérias , Infecções Bacterianas , Fungos , Sequenciamento de Nucleotídeos em Larga Escala , Metagenômica , Micoses , Humanos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Fungos/genética , Fungos/isolamento & purificação , Fungos/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/classificação , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia , Metagenômica/métodos , Micoses/diagnóstico , Micoses/microbiologia , Automação Laboratorial/métodos , Sensibilidade e Especificidade , Técnicas de Diagnóstico Molecular/métodos , Fatores de Tempo , Biologia Computacional/métodos , Masculino , Feminino , Criança , Adolescente , Adulto , Pré-Escolar
15.
Jpn J Ophthalmol ; 68(3): 233-242, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38658453

RESUMO

PURPOSE: To evaluate and compare the results of the conjunctival flap (CF) and cryopreserved amniotic membrane graft (AMG) in the management of fungal corneal ulcers either with complications or non-responsive to medical treatment. STUDY DESIGN: A retrospective observational study. METHODS: Medical records of 30 patients with culture-positive fungal corneal ulcer treated with either CF or AMG (15 eyes in each group) in real world settings were retrieved for analysis. After the surgical procedure, patients were followed up on days 1, 7, 14, 21, 30, 60, 90, 120, and 180 to explore the outcomes of the operations along with complications. RESULTS: Infecting fungi were of genus Fusarium (n = 11), Aspergillus (n = 10), Mucor (n = 4) and Penicillium (n = 10). The most common indication was resistant ulcer with perforation. After the procedure, epithelization was completed in 11(73.33%) patients in the CF, and 13 patients in the (86.67%) AMG group. Visual acuity improvement was significantly better in the latter group (CF: 1 [6.67%] vs. AMG: 7 [46.67%], p = 0.023). Flap failure occurred in 4 patients (26.67%) from the CF and 2 (13.33%) from the AMG group. No significant differences were found between the two groups regarding success rate (p = 0.651), epithelialization time (p = 0.691), healing of corneal ulcer (p = 0.651), and postoperative stability (p = 0.651) of the flaps. CONCLUSIONS: CF and AMG are both effective for the management of refractory fungal corneal ulcers. However, AMG appears to improve visual acuity better than CF.


Assuntos
Âmnio , Túnica Conjuntiva , Úlcera da Córnea , Infecções Oculares Fúngicas , Retalhos Cirúrgicos , Acuidade Visual , Humanos , Úlcera da Córnea/cirurgia , Úlcera da Córnea/microbiologia , Úlcera da Córnea/diagnóstico , Estudos Retrospectivos , Masculino , Âmnio/transplante , Feminino , Infecções Oculares Fúngicas/microbiologia , Infecções Oculares Fúngicas/cirurgia , Infecções Oculares Fúngicas/diagnóstico , Pessoa de Meia-Idade , Túnica Conjuntiva/microbiologia , Túnica Conjuntiva/cirurgia , Adulto , Idoso , Resultado do Tratamento , Seguimentos , Fungos/isolamento & purificação
16.
Microbiol Spectr ; 12(5): e0324523, 2024 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-38602397

RESUMO

Microorganisms are a crucial component of lake ecosystems and significant contributors to biogeochemical cycles. However, the understanding of how primary microorganism groups (e.g., bacteria and fungi) are distributed and constructed within different lake habitats is lacking. We investigated the bacterial and fungal communities of Hulun Lake using high-throughput sequencing techniques targeting 16S rRNA and Internal Transcribed Spacer 2 genes, including a range of ecological and statistical methodologies. Our findings reveal that environmental factors have high spatial and temporal variability. The composition and community structures vary significantly depending on differences in habitats. Variance partitioning analysis showed that environmental and geographical factors accounted for <20% of the community variation. Canonical correlation analysis showed that among the environmental factors, temperature, pH, and dissolved oxygen had strong control over microbial communities. However, the microbial communities (bacterial and fungal) were primarily controlled by the dispersal limitations of stochastic processes. This study offers fresh perspectives regarding the maintenance mechanism of bacterial and fungal biodiversity in lake ecosystems, especially regarding the responses of microbial communities under identical environmental stress.IMPORTANCELake ecosystems are an important part of the freshwater ecosystem. Lake microorganisms play an important role in material circulation and energy flow owing to their unique enzymatic and metabolic capacity. In this study, we observed that bacterial and fungal communities varied widely in the water and sediments of Hulun Lake. The primary factor affecting their formation was identified as dispersal limitation during stochastic processes. Environmental and geographical factors accounted for <20% of the variation in bacterial and fungal communities, with pH, temperature, and dissolved oxygen being important environmental factors. Our findings provide new insights into the responses of bacteria and fungi to the environment, shed light on the ecological processes of community building, and deepen our understanding of lake ecosystems. The results of this study provide a reference for lake management and conservation, particularly with respect to monitoring and understanding microbial communities in response to environmental changes.


Assuntos
Bactérias , Biodiversidade , Fungos , Lagos , Microbiota , Lagos/microbiologia , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Bactérias/genética , Bactérias/classificação , Bactérias/isolamento & purificação , Microbiota/genética , Ecossistema , RNA Ribossômico 16S/genética , Processos Estocásticos , Micobioma , Temperatura , Concentração de Íons de Hidrogênio , China
17.
mSystems ; 9(5): e0000424, 2024 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-38591897

RESUMO

Seed endophytic microbiomes are shaped by host and environmental factors and play a crucial role in their host growth and health. Studies have demonstrated that host genotype, including hybridization, affects seed microbiomes. Heterosis features are also observed in root-associated microbiomes. It remains unclear, however, whether heterosis exists in seed endophytic microbiomes and whether hybrid microbiota provide noticeable advantages to host plant growth, especially to seed germination. Here, we investigated the structure of seed endophytic bacterial and fungal communities from three hybrid rice varieties and their respective parents using amplicon sequencing targeting 16S rRNA and ITS2 genes. Heterosis was found in diversity and composition of seed endophytic microbiomes in hybrids, which hosted more diverse communities and significantly higher abundances of plant growth-promoting taxa, such as Pseudomonas and Rhizobium genera compared with their parental lines. Co-occurrence network analysis revealed that there are potentially tighter microbial interactions in the hybrid seeds compared with their parent seeds. Finally, inoculation of seed-cultivable endophytes, isolated from hybrids, resulted in a greater promotion of seed germination compared with those isolated from parent lines. These findings suggest that heterosis exists not only in plant traits but also in seed endophytic microbiota, the latter in turn promotes seed germination, which offers valuable guidance for microbiome-assisted rice breeding.IMPORTANCEGenetic and physiological changes associated with plant hybridization have been studied for many crop species. Still, little is known about the impact of hybridization on the seed microbiota. In this study, we indicate that hybridization has a significant impact on the endophytic bacterial and fungal communities in rice seeds. The seed endophytic microbiomes of hybrids displayed distinct characteristics from those of their parental lines and exhibited potential heterosis features. Furthermore, the inoculation of seed-cultivable endophytes isolated from hybrids exhibited a greater promotion effect on seed germination compared with those isolated from the parents. Our findings make a valuable contribution to the emerging field of microbiome-assisted plant breeding, highlighting the potential for a targeted approach that aims to achieve not only desired plant traits but also plant-beneficial microbial communities on the seeds.


Assuntos
Endófitos , Germinação , Vigor Híbrido , Microbiota , Oryza , Sementes , Oryza/microbiologia , Oryza/genética , Oryza/crescimento & desenvolvimento , Endófitos/genética , Sementes/microbiologia , Sementes/genética , Sementes/crescimento & desenvolvimento , Vigor Híbrido/genética , Microbiota/genética , Hibridização Genética , RNA Ribossômico 16S/genética , Bactérias/genética , Bactérias/classificação , Bactérias/isolamento & purificação , Fungos/genética , Fungos/isolamento & purificação , Fungos/classificação
18.
World J Microbiol Biotechnol ; 40(6): 175, 2024 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-38647735

RESUMO

The demand for environment-friendly cleanup techniques has arisen due to an increase in environmental pollutants. Fungi is the most prevalent and effective class of heavy metal-resistant microorganisms with the ability to leach metals. The objective of the present study was to isolate the fungi from the agricultural soil of Kashmir valley, investigate their multi-metal tolerance to heavy metals and evaluate the metal uptake capacities of the resistant fungi. The fungi were isolated and identified on the basis of morphological and molecular approach (ITS1 and ITS4). The tolerance limits of the isolated fungal strains to various doses of lead (Pb), cadmium (Cd), zinc (Zn), chromium (Cr), copper (Cu), nickel (Ni), and cobalt (Co) was evaluated. Five fungal strains, Aspergillus niger, Fusarium oxysporum, Fusarium verticillioides, Aspergillus fischeri, Epicoccum mackenziei were isolated from the soil samples. To the best of our knowledge, this is the first report on the study of metal resistance of Aspergillus fischeri and Epicoccum mackenziei. Among the identified fungal species, Aspergillus niger and Fusarium oxysporum were found to be most tolerant with a minimum inhibitory concentration (MIC) of 600 ppm against Cu and Cr respectively. Results indicated removal of considerable amount of heavy metals by some of the fungi. The highest metal uptake of 8.31 mg/g was found in Fusarium verticillioides for Zn. Surprisingly, these fungal strains demonstrated resistance to metal concentrations above the levels that are universally acceptable for polluted soils, and hence prove to be appealing contenders for use as bioremediation agents for cleaning up heavy metal-polluted environments.


Assuntos
Fungos , Fusarium , Metais Pesados , Testes de Sensibilidade Microbiana , Microbiologia do Solo , Poluentes do Solo , Metais Pesados/metabolismo , Poluentes do Solo/metabolismo , Fungos/efeitos dos fármacos , Fungos/isolamento & purificação , Fungos/classificação , Fungos/metabolismo , Fusarium/isolamento & purificação , Fusarium/efeitos dos fármacos , Fusarium/metabolismo , Biodegradação Ambiental , Aspergillus niger/isolamento & purificação , Aspergillus niger/efeitos dos fármacos , Aspergillus niger/metabolismo , Solo/química , Aspergillus/efeitos dos fármacos , Aspergillus/metabolismo , Aspergillus/isolamento & purificação
19.
World J Microbiol Biotechnol ; 40(6): 185, 2024 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-38683236

RESUMO

Stalk rot disease is a major constraint in maize production and till date reported to be caused by two to three species of phytopathogenic fungi but, in our present study, we disclose the first report of stalk rot is caused by complex species of phytopathogens, which belongs to five different genera. Therefore, to substantiate these findings, a total of 105 diseased samples of maize were collected from 21 different locations in six different geographical locations of India from which 48 isolates were used for the research study. Morphological features such as pigmentation, colony color, type of mycelium and pattern of mycelium was examined using macro and microscopic methods. A total of 11 different spp. of pathogens belonging to the five different genera: Fusarium verticillioides (56.25%), F. equiseti (14.5%), F. andiyazi (6.25%), F. solani (2.08%), F. proliferatum (2.08%), F. incarnatum (2.08%), Lasidioplodia theobrame (6.25%), Exserohilum rostrtum (4.16%), Nigrospora spp. (4.16%). and Schizophyllum commune (2.08%) were identified by different housekeeping genes (ITS, TEF-1α, RPB2 and Actin). Fusarium verticillioides, F. equiseti and F. andiyazi were major pathogens involved in stalk rot. This is the first report on F. proliferatum, F. solani, F. incarnatum, Lasidioplodia theobrame, Exserohilum rostrtum, Nigrospora spp. and Schizophyllum commune causing stalk rot of maize and their distribution in the different states of India. Studies on population dynamics of PFSR will enhance the understanding of pathogen behavior, virulence, or its association with different pathogens across India, which will facilitate the development of resistant maize genotypes against the PFSR.


Assuntos
Fusarium , Filogenia , Doenças das Plantas , Zea mays , Zea mays/microbiologia , Doenças das Plantas/microbiologia , Índia , Fusarium/genética , Fusarium/classificação , Fusarium/isolamento & purificação , Fusarium/patogenicidade , DNA Fúngico/genética , Fungos/genética , Fungos/classificação , Fungos/isolamento & purificação , Fungos/patogenicidade , Variação Genética
20.
Sci Rep ; 14(1): 9318, 2024 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-38654024

RESUMO

Endophytes of Panax have the potential to produce their host plant secondary metabolites, ginsenosides. Panax sokpayensis, an endemic traditional medicinal plant of the Sikkim Himalayas was explored for the isolation of endophytic fungi. In the present study, we have isolated 35 endophytic fungal cultures from the rhizome of P. sokpayensis and screened for ginsenosides production by HPLC by comparing the peak retention time with that of standard ginsenosides. The HPLC analysis revealed that out of 35 isolates, the mycelial extracts of four fungal endophytes (PSRF52, PSRF53, PSRF49 and PSRF58) exhibited peaks with a similar retention time of the standard ginsenoside, Compound K (CK). LC-ESI-MS/MS analysis led to the confirmation of ginsenoside CK production by the four fungal endophytes which showed a compound with m/z 639.6278, similar to that of standard ginsenoside CK with yield in potato dextrose broth flask fermentation ranging from 0.0019 to 0.0386 mg/g of mycelial mass in dry weight basis. The four prospective fungal endophyte isolates were identified as Thermothielavioides terrestris PSRF52, Aspergillus sp. PSRF49, Rutstroemiaceae sp. strain PSRF53, and Phaeosphaeriaceae sp. strain PSRF58 based on ITS sequencing. The present finding highlights the need for further study on growth optimization and other culture parameters to exploit the endophytes as an alternative source for ginsenoside CK production.


Assuntos
Endófitos , Fermentação , Ginsenosídeos , Panax , Ginsenosídeos/metabolismo , Endófitos/metabolismo , Endófitos/isolamento & purificação , Panax/microbiologia , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas em Tandem , Fungos/metabolismo , Fungos/isolamento & purificação , Rizoma/microbiologia
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